Abstract:
:Neuroscience relies on techniques for imaging the structure and dynamics of neural circuits, but the cell bodies of individual neurons are often obscured by overlapping fluorescence from axons and dendrites in surrounding neuropil. Here, we describe two strategies for using the ribosome to restrict the expression of fluorescent proteins to the neuronal soma. We show first that a ribosome-tethered nanobody can be used to trap GFP in the cell body, thereby enabling direct visualization of previously undetectable GFP fluorescence. We then design a ribosome-tethered GCaMP for imaging calcium dynamics. We show that this reporter faithfully tracks somatic calcium dynamics in the mouse brain while eliminating cross-talk between neurons caused by contaminating neuropil. In worms, this reporter enables whole-brain imaging with faster kinetics and brighter fluorescence than commonly used nuclear GCaMPs. These two approaches provide a general way to enhance the specificity of imaging in neurobiology.
journal_name
Neuronjournal_title
Neuronauthors
Chen Y,Jang H,Spratt PWE,Kosar S,Taylor DE,Essner RA,Bai L,Leib DE,Kuo TW,Lin YC,Patel M,Subkhangulova A,Kato S,Feinberg EH,Bender KJ,Knight ZA,Garrison JLdoi
10.1016/j.neuron.2020.05.005subject
Has Abstractpub_date
2020-08-05 00:00:00pages
454-469.e6issue
3eissn
0896-6273issn
1097-4199pii
S0896-6273(20)30351-2journal_volume
107pub_type
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